LOCUS       sequence-333335-        4111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for mammalian expression using the 
            human Ubiquitin C promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 4111)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C413-E33: attB4-UbCp>-attB1r
FEATURES             Location/Qualifiers
     source          1..4111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..51
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     43..60
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     44..60
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    116..211
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        211..1422
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1259..1278
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    1424..1547
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1619..1638)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1620..1638)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1643..1659)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1643..1659)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2084..2875
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2968..3556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3457..3476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3710..3727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3886..3913)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4005..4091)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
       61 cttaaaattc gagctcggtc cgatcgggcc cgcgttaacg ctaccatgga gctccaaata
      121 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt
      181 tttataatgc caactttgta tagaaaagtt ggcctccgcg ccgggttttg gcgcctcccg
      241 cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc
      301 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
      361 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
      421 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
      481 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
      541 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
      601 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
      661 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
      721 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
      781 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
      841 ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg
      901 agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
      961 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     1021 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     1081 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     1141 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     1201 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     1261 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     1321 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa
     1381 ttgtccgcta aattctggcc gtttttggct tttttgttag acgcaacttt gtacaaaaaa
     1441 gttgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     1501 aaacagacta cataatactg taaaacacaa catatgcagt cactatgaac caactactta
     1561 gatggtatta gtgacctgta gaattcgagc tctagagctg cagggcggcc gcgatatccc
     1621 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1681 aaatctctga tgttacattg cacaagatcg gtacccgggg atcctctaga ccagccagga
     1741 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     1801 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     1861 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     1921 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     1981 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     2041 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
     2101 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     2161 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     2221 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     2281 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     2341 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     2401 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     2461 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     2521 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     2581 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     2641 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     2701 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     2761 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     2821 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     2881 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2941 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3001 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3061 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3541 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3601 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3661 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3721 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3781 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3841 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     3901 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     3961 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     4021 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
     4081 cgttttattt gatgcctggc agttccctac t
//
